MHC class 1 detection by Immunocytochemistry - (May/25/2011 )

Have to detect MHC class 1 expression through Immunocytochemistry.Looking for good protocol.Any experience would be helpful

-santhi-

use Abcam protocol for ICC

-Curtis-

Santhi

1. What type of cells are you using? Mouse, human?

2. Is there a specific MHC class I molecule you are looking for?

3. What fluorescence channels will you be able to pick up on your confocal? Do you have a preferred fluorophore?

4. Are you going to label your antibodies yourself? Buy prelabelled? Use a secondary?

5. Do you want to visualise the MHC I on the cell surface? Internally also?

If you tell us more about what you are doing and why, we might be able to help you more?
Your first step though, should be to go and do some research on immunohistochemistry so you know what you are actually doing.

-leelee-

Hi Santhi, sorry if I came over a little harsh, I just re-read my post and it was a little blunt!
I've done labelling for both H2kk and H2kd for flow cytometry before, but not immunohistochem.

If you want to have a think and a look around at some protocols (and I will too when I have some time), then post up what you've got and we'll figure it out.

Do you happen to know what mouse cells you have (just to double check you have the right type for the molecules you are trying to detect).

Also, do you want to measure both molecules in the same sample, or do them seperately?